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目的 观察微小RNA-132(miR-132)转染人胰腺癌细胞株SW1990后对细胞增殖及凋亡的影响,并探讨其作用机制.方法 采用实时荧光定量PCR (RT-qPCR)法检测28例胰腺癌及匹配的癌旁正常胰腺组织miR-132的表达.采用脂质体将miR-132转染SW1990细胞,以未转染及转染错义miR-132的细胞分别作为空白对照和阴性对照.应用CCK-8法、DAPI染色法检测细胞的增殖及凋亡;将转染细胞种植于裸鼠皮下成瘤,应用TUNEL法检测种植瘤细胞凋亡;免疫组化法检测转染细胞的mucin-4、HER-2、p-FAK蛋白表达及种植瘤组织mucin-4、Ki-67蛋白表达.结果 28例胰腺癌及癌旁正常胰腺组织miR-132的相对表达量分别为0.46±0.11和1.24±0.36,差异有统计学意义(P＜0.05).转染细胞的miR-132表达量为2.95 ±0.46,显著高于阴性对照组的0.84±0.17(P ＜0.05);转染组细胞培养48、72、96 h时的存活率分别为阴性对照组56.5％、44.7％、37.4％(P值均＜0.05);细胞凋亡率为41.6％,显著高于阴性对照组的5.7％(P＜0.05);转染细胞mucin-4、HER-2、p-FAK蛋白的表达较阴性对照组显著下调(0.76 ±0.14比2.94±0.42,0.34±0.04比1.75±0.33,0.27±0.03比2.74±0.24,P值均＜0.01).与阴性对照组比较,转染组裸鼠皮下移植瘤生长明显被抑制[(0.23±0.05)g比(0.59±0.13)g,P＜0.05],瘤内肿瘤细胞凋亡明显增加[(21.7±4.7)％比(5.2±0.7)％,P＜0.05],mucin-4和Ki-67蛋白表达显著下调(64.35±7.16比281.34±36.62,30.75±4.61比148.05±21.34,P值均＜0.01).而阴性对照组与空白对照组间的差异均无统计学意义.结论 转染miR-132对胰腺癌SW1990细胞有显著的增殖抑制和凋亡诱导作用,其机制可能与下调mucin-4、HER-2、p-FAK等蛋白的表达有关.

Objective To observe the effect of miR-132 transfection on proliferation and apoptosis of pancreatic cancer SW1990,and to explore the underlying mechanism.Methods The expression of miR-132 in the pancreatic cancer tissue and the adjacent tissues in 28 pancreatic cancer patients were detected by realtime quantitative polymerase chain reaction (RT-qPCR).miR-132 was transfected into SW1990 cells by using liposome method,untransfected cells and cells with missense miR-132 transfection were used as black control and negative control.The proliferation and apoptosis was detected by CCK-8 assay and DAPI staining.The transfected cells were implanted in nude mice as xenograft tumor,and TUNEL was used to detect the apoptosis; immunohistochemistry was used to detect the expression of Ki-67 and mucin-4 protein in the xenograft tumors and mucin-4,HER-2,p-FAK protein in transfected cells.Results The expression levels of miR-132 in pancreatic cancer tissue and adjacent tissues were 0.46 ± 0.11 and 1.24 ± 0.36,and the difference between the two groups was statistically significant (P ＜ 0.05).The expression level of miR-132 in transfected SW1990 cells was 2.95 ± 0.46,which were significantly higher than those in negative control (0.84 ± 0.17) ; the survival rate of transfected cells at 48,72,96 h was 56.5％,44.7％,37.4％ of negative control cells.The apoptosis rate in transfected cells was 41.6％,and the corresponding value was 5.7％ in negative control,and the difference was statistically significant (P ＜ 0.05).The expression levels of mucin-4,HER-2,p-FAK in nagative control were 2.94 ± 0.42,1.75 ± 0.37 and 2.74 ± 0.24,and the corresponding values in transfected cells were 0.76 ± 0.14,0.34 ± 0.04 and 0.27 ± 0.03,and the difference between the two groups was statistically significant (P ＜0.05).In vivo,the growth of xenograft tumors in transfected nude mice was significantly inhibited [(0.23 ± 0.05) vs (0.59 ± 0.13) g,P ＜ 0.05],the apoptosis of xenograft tumor cells was significant increased [(21.7 ± 4.7) ％ vs (5.2 ± 0.7) ％,P ＜0.05].The expressions of mucin-4 and Ki-67 protein in nagative control was 281.34 ± 36.62 and 148.05 ± 21.34,and the corresponding values in transfection group were 64.35 ± 7.16 and 30.75 ± 4.61,and the difference was statistically significant (P ＜0.05).Conclusions miR-132 transfection has an effect of inhibiting proliferation and promoting apoptosis on SW1990 cells,and the mechanism may be down-regulation of mucin-4,HER-2,p-FAK protein rxpression.